Prostaglandin A1 triggers Mayaro virus inhibition and heat shock protein 70 expression in an epithelial cell model

Abstract INTRODUCTION: The Mayaro virus (MAYV), which is an arbovirus closely related to the Chikungunya virus, causes a dengue-like acute illness that is endemic to Central and South America. We investigated the anti-MAYV activity of prostaglandin A1 (PGA1), a hormone which exhibits antiviral activity against both ribonucleic acid (RNA) and deoxyribonucleic acid (DNA) viruses. Further, we examined the effects of inducting the stress protein HSP70 following PGA1 treatment. METHODS: Hep-2 cells infected with MAYV were treated with PGA1 (0.1-6μg/ml) 12h before infection and for different periods post-infection. Inhibition of viral replication inhibition was analyzed via viral titer determination, whereas the effect of PGA1 on viral morphogenesis was examined via transmission electron microscopy (TEM). Autoradiography (with 35S methionine labeling) and western blotting were used to assess the effect of PGA1 treatment on viral and cellular protein synthesis, and on HSP70 induction, respectively. RESULTS: PGA1 strongly reduced viral replication in Hep-2 cells, particularly when added during the early stages of viral replication. Although PGA1 treatment inhibited viral replication by 95% at 24 hours post-infection (hpi), viral structural protein synthesis was inhibited only by 15%. TEM analysis suggested that PGA1 inhibited replication before viral morphogenesis. Western blot and densitometry analyses showed that PGA1 treatment increased HSP70 protein levels, although this was not detectable via autoradiography. CONCLUSIONS: PGA1 inhibits MAYV replication in Hep-2 cells at early stages of viral replication, prior to production of viral structural proteins, possibly via HSP70 induction.

Ultrastructure of the secretory epithelial cells of the bulbo-urethral glands (Cowper's gland) in the Indian fruit bat, Rousettus leschenaulti (Desmarest) during the reproductive cycle / Ultraestructura de las células epiteliales secretoras de las glándulas bulbouretrales (glándula de Cowper) en el murciélago de la India, Rousettus leschenaulti (Desmarest) durante el ciclo reproductivo

The present paper describes the ultrastructural characteristics of the bulbo-urethral gland (Cowper' glands) of the Indian fruit bat, R. leschenaulti during sexually inactive-breeding cycle. Cyclic histological changes during the seasonal breeding quiescence cycle are not well marked. There are no marked differences. The ultrastructural characteristic of the secretory epithelial cells of Bulbo-Uretrhal Gland gland have been studied during different phases of reproductive cycle. The secretory epithelial cells are characterized by the well-developed rough endoplasmic reticulum, extensive developed complexus golgiensis (Golgi apparatus) and mitochondria. Mitochondria with lamellate cristae are dispersed in the cytoplasm. Three different types of secretory granules can be identified on the basis of electron density. These granules are not of different types but they represent the different stages of granule maturation. The secretory products of bulbo-urethral gland of bat are released into lumen both by apocrine and merocrine modes. The functional significance of the secretions of the bulbo-urethral glands in reproduction is discussed.

El presente trabajo describe las características ultraestructurales de las glándulas bulbouretrales (glándulas de Cowper) del murciélago de la fruta de la India, R. leschenaulti durante el ciclo inactivo de reproducción sexual. Los cambios histológicos cíclicos durante el ciclo de quiescencia estacional de la cría no están bien determinados. No hay diferencias marcadas. La característica ultra estructural de las células epiteliales secretoras de la glándula bulbouretral ha sido estudiada durante diferentes fases del ciclo reproductivo. Las células epiteliales secretoras se caracterizan por un retículo endoplasmático rugoso bien desarrollado, el complexus golgiensis (complejo de Golgi) y mitocondrias desarrollados extensamente. Las mitocondrias con crestas lamelares se encontraron dispersas en el citoplasma. Se pueden identificar tres tipos diferentes de gránulos secretores en base a la densidad de electrones. Estos gránulos no son de tipos diferentes, sino que representan las diferentes etapas de maduración del gránulo. Los productos secretores de las glándulas bulbouretrales de murciélagos son liberados en el lumen tanto por modos apócrinos como merócrinos. Se discute la importancia funcional de las secreciones de la glándula bulbouretral en la reproducción.

Poros Nucleares y Función Celular en Epitelio Mamario / Nuclear Pores and Cellular Function

El presente artículo tiene como objetivo central evidenciar la interesante relación que se establece entre la función celular y el número de poros nucleares, relación que modula el activo intercambio nucleo-citoplasmatico en distintas etapas del ciclo celular de la estirpe HC11.

The main objective of this article is related to the study of different existing relationships between cellular function and the number of nuclear pores in order to explain the amount of nuclear-cytoplasmatic exchange through HC11 cell cycle stages.

Structural modifications of the prostate in hypoxia, oxidative stress, and chronic ischemia

PURPOSE: Clinical studies have reported a correlation between pelvic ischemia and voiding dysfunction in elderly men. The aim of this study was to identify and compare prostate structural modifications in cultured cells and in a rabbit model after exposure to hypoxia, oxidative stress, and chronic ischemia. MATERIALS AND METHODS: Cultured human prostate smooth muscle cells (SMCs), epithelial cells (ECs), and stromal cells (SCs) were incubated under normoxia, hypoxia, and oxidative stress conditions by use of a computerized oxycycler system. We developed a rabbit model of chronic prostate ischemia by creating aorto-iliac arterial atherosclerosis. Markers of oxidative stress were examined by using fluorometric analysis and enzyme immunoassay. Prostate structure was examined by using Masson's trichrome staining and transmission electron microscopy (TEM). RESULTS: Lipid peroxidation was found in SMCs exposed to hypoxia and in all cell types exposed to oxidative stress. We identified protein oxidation in ECs exposed to hypoxia and in all cell types exposed to oxidative stress. Markers indicating oxidative damage were present in chronically ischemic rabbit prostate tissue. These reactions were associated with DNA damage. Prostate ischemia resulted in epithelial atrophy, loss of smooth muscle, and diffuse fibrosis. TEM showed swollen mitochondria with degraded cristae, loss of membrane, loss of Golgi bodies, degenerated nerves, and disrupted cell-to-cell junctions. CONCLUSIONS: Human prostate cells exhibited differential reactions to hypoxia and oxidative stress with widespread DNA damage. Structural modifications in ischemic prostate tissue were similar to those in cells exposed to oxidative stress. Structural changes due to ischemia and oxidative stress may contribute to prostatic noncompliance in aging men.

Human sepsis-associated Escherichia coli (SEPEC) is able to adhere to and invade kidney epithelial cells in culture

The adhesins of extraintestinal pathogenic Escherichia coli are essential for mediating direct interactions between the microbes and the host cell surfaces that they infect. Using fluorescence microscopy and gentamycin protection assays, we observed that 49 sepsis-associated E. coli (SEPEC) strains isolated from human adults adhered to and invaded Vero cells in the presence of D-mannose (100%). In addition, bacteria concentrations of approximately 2 x 10(7) CFU/mL were recovered from Vero cells following an invasion assay. Furthermore, PCR analysis of adhesin genes showed that 98.0% of these SEPEC strains tested positive for fimH, 69.4% for flu, 53.1% for csgA, 38.8% for mat, and 32.7% for iha. Analysis of the invasin genes showed that 16.3% of the SEPEC strains were positive for tia, 12.3% for gimB, and 10.2% for ibeA. Therefore, these data suggest that SEPEC adhesion to cell surfaces occurs through non-fimH mechanisms. Scanning electron microscopy showed the formation of microcolonies on the Vero cell surface. SEPEC invasiveness was also confirmed by the presence of intracellular bacteria, and ultrastructural analysis using electron transmission microscopy revealed bacteria inside the Vero cells. Taken together, these results demonstrate that these SEPEC strains had the ability to adhere to and invade Vero cells. Moreover, these data support the theory that renal cells may be the predominant pathway through which SEPEC enters human blood vessels.

Estandarización en la observación y clasificación de lesiones epiteliales premalignas y malignas / Standardization in the observation and classification of premalignant and malignant epithelial lesions

Los criterios histológicos para determinar el grado de displasia, la clasificación de Broders y el frente de invasión tumoral (FIT) son parámetros subjetivos no cuantificables que pueden indicar el grado de evolución de displasias y carcinomas. Un factor importante a considerar durante la valoración histológica, es la variabilidad del diagnóstico entre patólogos. El objetivo es estandarizar los criterios y determinar la variabilidad intra e inter observador en el diagnóstico de displasias y COCE. Se seleccionaron y estandarizaron los criterios morfológicos para el diagnóstico y se revisaron los casos seleccionados aleatoriamente por tres patólogos bucales (30 displasias y 30 carcinomas) del Laboratorio de Patología Clínica y Experimental de la DEPeI de la FO, UNAM. Cada patólogo analizó y registró los parámetros establecidos para displasia, COCE y FIT en 2 ocasiones. Se aplicó el test Kappa para valorar la concordancia intra e inter observador. El Observador 1 v/s el 2 obtuvo una concordancia para COCE de 0,75 y en displasias de 0,60 e intraobservador de 0,90. El observador 2 v/s el 3 presentó una concordancia para COCE de 0,75 y en displasias de 0,59 e intraobservador de 0,91. El Observador 3 Vs el 1 tuvo una concordancia para COCE de 0,77, y en displasias de 0,59 e intraobservador de 0,92. La concordancia intraobservador e interobservador en COCE fue de buena a excelente, pero en displasias fue aceptable confirmando que su evaluación presenta mayor grado de dificultad. Con una adecuada estandarización se puede obtener una buena concordancia entre patólogos.

In the histological criteria for determining the degree of dysplasia, the Broders classification and the front of tumor invasion (FTI) are unquantifiable subjective parameters that may indicate the degree of development of carcinomas. An important factor to consider during the histological evaluation is the variability in the diagnosis of pathologists. The objective to standardize criteria and determine the intra and inter-observer variability in the diagnosis of dysplasias and OSCC. We selected and standardized morphological criteria for the diagnosis, and the cases were reviewed randomly by three oral pathologists (30 dysplasias and 30 carcinomas) from the Laboratory of Clinical and Experimental Pathology of the FO DEPeI, UNAM. Each pathologist analyzed and recorded the parameters for dysplasia and OSCC FIT on two occasions. Kappa test was applied to assess intra and inter-observer agreement. Observer 1 v/s 2 match for OSCC was 0.75, 0.60 for dysplasias and intra observer 0.90. Observer 2 v/s 3 presented a concordance of 0.75 for OSCC, 0.59 for dysplasias and intra-observer 0.91. Observer 3 v/s observer 1 for OSCC was 0.77, 0.59 for dysplasias and intra-observer 0.92. Intra observer and inter-observer concordance in OSCC were good or excellent, but in dysplasia was acceptable, confirming that its assessment showed the greatest difficulty with proper standardization we can obtain a better consensus between pathologists.

El factor de crecimiento epidérmico y la diferenciación celular del epitelio mamario / Epidermal growth factor and mammary epithelial differentiation

El objetivo de este artículo es presentar una revisión relativa a evidenciar las características generales, estructura y funcionalidad de los factores de crecimiento con especial énfasis en precisar el rol que ejerce el factor de crecimiento epidérmico (EGF) como agente generador del proceso de diferenciación celular en el epitelio mamario.

The objective of this article is to present a review referred to general characteristics, such as structure and functionality of the growth factors, particularly those related to the Epidermal Growth Factor (EGF), as a responsible generator agent of cell differentiation at the mammary epithelial level.

Variaciones nucleolares en la transformación celular del epitelio mamario / Nucleolar variations in the cell transformation of mammary epithelium

Antecedentes: En la medida que las células pertenecientes a la línea HC11 son transfectadas con el onco-gén ras, asumen distintas propiedades resultando en tipos celulares transformados, modificando tanto sus componentes como sus funciones celulares, los cuales pueden ser cuantificadas mediante técnicas morfométricas. Objetivo: Evidenciar en términos cuantitativos y morfológicos las variaciones experimentadas por los nucleolos pertenecientes a células mamarias de la línea HC11 con el decorrer del mecanismo de transformación celular. Método: Se estudió a nivel de la microscopia electrónica de transmisión los tipos celulares en proceso de transformación (Q6 GM), en comparación con células francamente neoplásicas (Q6 IM), cuantificando variaciones de los nucleolos y su relación con estructuras involucradas en síntesis proteica. Resultados: Se evidencian diferencias estadísticamente significativas en el área, volumen y longitud entre los nucleolos pertenecientes a estos tipos celulares. Conclusión: Las células del epitelio mamario en proceso neoplásico presentan un notable aumento de sus nucleolos y sus ribonucleoproteínas, constituyentes que generarán básicamente ribosomas libres, que sintetizarán proteínas para ser utilizadas en el decorrer de las mitosis sucesivas y desreguladas.

Background: As cells belonging to the HC11 line become transfected with the ras oncogene, they assume different properties resulting in transformed cell types, with modified cell components and functions. These may be quantified by morphometric techniques. Objective: To provide quantitative and morphological evidence of the variations occurring in the nucleoles of HC11 line mammary cells as the cell transformation mechanism takes its course. Method: Transmission electron microscopy was used to study cell types in the transformation phase (Q6 GM) as compared with frankly neoplastic cells (Q6 IM), quantifying the variations between the nucleoles and their relation to structures involved in protein synthesis. Results: Statistically significant differences were found between the nucleoles belonging to these cell types with respect to area, volume and length. Conclusion: The nucleoles of mammary epithelial cells in the process of neoplasia, present a notable increase, and their constituent ribonucleoproteins will basically generate free ribosomes, synthesising proteins available for use in successive, unregulated mitosis.

Cytogenetic aberrations in primary cell cultures of the ovarian surface epithelium / Aberraciones citogenéticas en cultivos primarios del epitelio superficial del ovario

Our objective was to determine the presence of chromosomal abnormalities in primary cultures of ovarian surface epithelial cells in women of different ages with no history of cancer. Throughout conventional cytogenetic techniques, we analyzed chromosome spreads of cultured ovarian epithelial cells from 10 donors who were 50 or more years old (B) and 16 controls between 20 and 49 years old (A), belonging to the mestizo population in Bogota DC, Colombia. Of the 26 cultures that were analyzed in passage 1, 61.5% had an abnormal chromosome complement (62.5% in A, and 60% in B). Abnormalities included polyploidies, endoduplications and monosomies. Deletions in chromosomes 3 and 11 were found in just one metaphase. None of the samples showed weaknesses or breakpoints. After transforming and applying the exact student’s t-test for variance heterogeneity, we found significant differences in the frequency of metaphases, that were higher in A than in B (p=0.05), and in the frequency of polyploidies, which were higher in B than in A (p=0.044). Through the application of the Mann-Whitney test, we determined that the frequency of endoduplications was higher in A than in B (p=0.126), without reaching significant differences. There were no significant differences in the frequency of monosomies. The level of significance was set at p £ 0.05. Taking into account that polyploidization is a marker of chromosomal instability and that the risk of cancer arising from the ovarian surface epithelium augments substantially after menopause, the increase in the frequency of age-associated polyploidies could be used as a predictor of ovarian cancer in women from an ethnically homogeneous population as the mestizo one in Bogota DC.

El objetivo del presente trabajo fue determinar la presencia de anormalidades cromosómicas en cultivos primarios de células del epitelio superficial ovárico en mujeres de diferentes edades, sin antecedentes de cáncer. Mediante técnicas de citogenética convencional fueron analizados extendidos de células epiteliales ováricas histológicamente normales, provenientes de cultivos primarios de 10 donantes de 50 o más años (B) y de 16 donantes entre 20 y 49 años que se utilizaron como grupo control (A), pertenecientes a la población mestiza de Bogotá DC, Colombia. De 26 cultivos examinados en pase 1, 61,5% presentó complemento cromosómico anormal, 62,5% en A y 60% en B. Las anomalías numéricas halladas, todas en mosaico, incluyeron poliploidías, endoduplicaciones y monosomías. En una única célula en metafase de un cultivo, se presentaron deleciones en los cromosomas 3 y 11. Ninguna muestra presentó fragilidades o roturas. Previa aplicación de transformaciones, con la prueba exacta t-student para varianzas heterogéneas, se encontraron diferencias significativas en la frecuencia de células con metafase normal, mayor en A que en B (p=0,05) y en la de poliploidías, mayor en B que en A (p=0,044). Con la prueba exacta de Mann-Whitney se determinó que la frecuencia de endoduplicaciones en A fue mayor que en B (p=0,126), sin alcanzar diferencias significativas y que no hubo diferencias significativas en la frecuencia de monosomías. El nivel de significación fue p £ 0,05. Si se tiene en cuenta que la poliploidización es un marcador de inestabilidad cromosómica y, que además, el riesgo de aparición de cáncer derivado del epitelio superficial del ovario aumenta sustancialmente después de la menopausia, el incremento en la frecuencia de poliploidías asociado con la edad podría ser utilizado como predictor de cáncer ovárico en mujeres de una población étnicamente homogénea como la población mestiza de Bogotá DC.

Cáncer de mama e índice de masa corporal: en mujeres posmenopáusicas / Breast cancer and body mass index: in posmenopausal women

El cáncer de mama representa la segunda causa de muerte por neoplasias malignas en mujeres venezolanas. Sobrepeso y obesidad son factores de riesgo de esta patología, por la producción de estrógenos en el tejido adiposo por procesos enzimáticos. Comprobar la relación entre carcinoma mamario e índice de masa corporal en mujeres posmenopáusicas. Estudio cuantitativo con nivel correlacional, tipo casos y controles. La muestra de tipo no probabilística estuvo compuesta por 103 mujeres posmenopáusicas con cáncer de mama (casos), y 100 mujeres posmenopáusicas sin carcinoma mamario (controles) que acudieron a consulta de la Unidad de Mastología del Centro Médico “Dr. Rafael Guerra Méndez” en Valencia, Estado Carabobo entre 1992 y 2007. La recolección de datos se hizo mediante revisión de historias clínicas, representándose los resultados en mediana, rango intercuartil, prueba de Chi cuadrado, Chi cuadrado con corrección de Yates y la prueba de Wilcoxon. Se clasificaron a las pacientes con y sin cáncer de mama según su índice de masa corporal, el 44,66 por ciento tenían sobrepeso y 36,9 por ciento obesidad; y en los controles el 38 por ciento presentó sobrepeso, 36 por ciento obesidad y 1 por ciento desnutrición. No se estableció una asociación estadísticamente significativa al 95 por ciento de confianza entre el índice y cáncer de mama. No existe relación entre cáncer de mama y el índice de masa corporal en la muestra estudiada. Se sugiere realizar estudios similares en muestras más significativas para así obtener resultados más fidedignos.

Breast cancer is currently the second leading cause of death from malignant tumours in women’s. Recent investigations classified overweight and obesity as risk factors for this disease, the association is due to the production of estrogens in fatty tissue by enzymatic processes. In these work we can determine the relationship between breast carcinoma and body mass index in postmenopausal women. A quantitative correlation study, type cases and controls. The non-probability sample consisted of 103 postmenopausal women with breast cancer (cases), 100 postmenopausal women without breast carcinoma (controls) who came to consult at hospital “Dr. Guerra Mendez” mastology unit, in Valencia, Carabobo between 1992 and 2007. Data collection was done through review of medical records, representing the results in median, interquartile range, Chi square test, Chi square with Yates’ correction and the Wilcoxon test. Patients were classifying in two groups, with and without breast cancer, according to body mass index, noting that in the cases the 44.66 percent were overweight and the 36.9 percent were obese. In the other hand the controls had 38 percent of overweight people, 36 percent obese and 1 percent malnutrition. There was no a statistically significant 95 percent confidence association between index and breast cancer. There is no relationship between breast cancer and the body mass index in the sample studied. These studied suggested to make similar projects with a more samples, and let us to get meaningful and more reliable results.

Mitochondria in the midgut epithelial cells of sugarcane borer parasitized by Cotesia flavipes (Cameron, 1891) / Mitocôndria nas Células Epiteliais do Intestino Médio da Broca-da-cana Parasitada por Cotesia flavipes

The sugarcane borer Diatraea saccharalis (Lepidoptera: Crambidae) has been controlled by Cotesia flavipes (Hymenoptera: Braconidae); however, very little is known about the effect of the parasitism in the host organs, including the midgut. This work aims to verify mitochondrial alteration in the different midgut epithelial cells of D. saccharalis parasitized by C. flavipes. Midgut fragments (anterior and posterior region) of both non-parasitized and parasitized larvae were processed for transmission electron microscopy. The mitochondria of midgut epithelial cell in the parasitized larvae exhibit morphological alteration, represented by matrix rarefaction and vacuolisation. These mitochondrial alterations are more pronounced in the anterior midgut region during the parasitism process, mainly in the columnar cell.

Diatraea saccharalis (Lepidoptera: Crambidae), broca da cana-de-açúcar, tem sido controlada por Cotesia flavipes (Hymenoptera: Braconidae); pouco se sabe sobre o efeito do parasitismo nos diferentes órgãos do inseto hospedeiro, principalmente no intestino médio. O objetivo desse trabalho foi verificar as alterações mitocondriais das diferentes células epiteliais do intestino médio de larvas de D. saccharalis parasitadas por C. flavipes. Fragmentos do intestino médio (regiões anterior e posterior) de larvas de D. saccharalis não-parasitadas e parasitadas foram processados para microscopia eletrônica de transmissão. As mitocôndrias das células epiteliais do intestino médio de larvas parasitadas exibem alterações, especialmente rarefação e vacuolização da matriz, que foram mais pronunciadas nas células epiteliais da região anterior do intestino médio na vigência do parasitismo, em especial nas células colunares.

Morfometría del nucleolo en la diferenciación celular del epitelio mamario / Nucleolus morphometry in the cellular differentiation of mammary epithelium

Antecedentes: El proceso biológico de diferenciación celular es la traducción de múltiples procesos nucleares y citoplasmáticos que determinan cambios complejos y fundamentales en la ultraestructura, bioquímica y fsiología celular, los cuales pueden ser cuantifcados mediante técnicas morfométricas. Objetivo: Evidenciar en términos cuantitativos y morfológicos las variaciones experimentadas por los nucleolos pertenecientes a células mamarias de la línea HC11 tanto normales como en mecanismo de diferenciación. Método: Se estudió a nivel de la microscopía electrónica de transmisión los tipos celulares en etapa de proliferación (HC11 GM) en comparación con células en estadio de diferenciación (HC11 IM), cuantifcando las variaciones de los nucleolos y su relación con estructuras involucradas en síntesis proteica. Resultados: Se evidencian diferencias estadísticamente signifcativas referentes al área, volumen y longitud entre los nucleolos pertenecientes al tipo celular normal-proliferante y el que se encuentra en proceso de diferenciación. Conclusión: Las células mamarias en proceso de diferenciación presentan una notable disminución de sus nucleolos, y sus ribonucleoproteínas constitutivas generarán básicamente ribosomas adheridos al retículo endoplasmático rugoso, sintetizando proteínas de exportación.

Background: The biological process of cell differentiation is the traslation of multiple nuclear and cytoplas-mic processes that determine complex and fundamental changes in ultrastructure, biochemistry and cell physiology, which can be quantifed using morphometric techniques. Objective: To show in quantitative and morphological terms changes experienced by the nucleolus belonging to HC11 line mammary cells both, in proliferating and differentiation process. Methods: A study at the level of transmission electron microscopy of cell types in stage of cell proliferation in comparison with stage of differentiation was designed to quantify variations of nucleolus and their relation to structures involved in protein synthesis. Results: Marked differences in the area, volume and length of the nucleolus were found between normal-proliferating cell types and those in mechanism of differentiation. Conclusion: The mammary cells in differentiation process show a dramatic decline in its nucleoli and their ribonucleoproteins generate basically ribosomes attached at endo-plasmic reticulum, synthesizing export proteins.

Morphological study of the cervical salivary glands in the nine-banded armadillo / Estudio morfológico de las glándulas salivales cervicales en el armadillo de nueve-bandas

The cervical salivary glands of the armadillo Dasypus novemcinctus was examined by light microscopy. These glands are situated on either side of the neck, divide in lobes and show a presence of a salivary bladder, associated with the main ducts of the gland. This gland is histologically a typical mixed glands, containing both mucous and serous elements, with mucous acini as the predominant secretory unit. The bladder itself is composed of a wall made up of pseudostratified epithelium, skeletal muscle and connective tissue. In general, the morphology of the cervical salivary glands appears similar to that described in other species of the mammals.

Las glándulas salivales cervicales del armadillo Dasypus novemcinctus fueron examinadas por microscopía de luz. Estas glándulas se encuentran a ambos lados del cuello, divididas en lóbulos y muestran la presencia de una vejiga salival, asociada con los principales conductos de la glándula. Esta glándula es histológicamente una típica glándula mixta, que contiene tanto elementos mucosos y serosos, con acinos mucosos como la principal unidad secretora. La vejiga en sí se compone de una pared formada por epitelio pseudoestratificado, músculo esquelético y tejido conectivo. En general, la morfología de las glándulas salivales cervicales parece similar a la descrita en otras especies de mamíferos.

Características regionais sobre a ultra-estrutura das células principais do epitélio do ducto epididimal de Agouti paca / Regional ultra structural features of principal cells from the epididymal epithelium lining of Agouti paca

As células principais (P) do epitélio de revestimento do epidídimo de paca foram relacionadas com processos citofisiológicos de endocitoses do tipo adsortiva e de fase fluida, respectivamente, aparentemente realizando também secreção apócrina. Essas funções foram propostas embasando-se em características de ultra-estrutura das células P, em cujos citoplasma observaram-se um expressivo número de vesículas, com diferentes formas, tamanhos e presença de conteúdo internalizado em algumas das vesículas revestidas por endomembranas, ocorrendo ainda caveolas e vesículas diminutas localizadas junto à borda apical de microvilos. Ademais, observaram-se vesículas grandes e revestidas e/ou com superfícies lisas; endossomos, e lisossomos de localização predominantemente apical. Uma via de secreção apócrina foi sugerida com base na ocorrência de expansões (protrusões), citoplasmáticas intraluminais nas células P.

The principal (P) cells of epididymidis surface epithelium of Agouti paca were related to processes of adsorptive endocytosis and phase-fluid endocytosis, as well as protein secretion apparently also occur.These findings had been proposed on the base the cytoplasmic ultrastructural features of P cells in which were seen an expressive number of vesicles with several shapes, sizes and internalized content occurring also smaller pits and pale small vesicles located next to the apical brush border of microvilli. Moreover, occurred coated vesicles, smooth surface vesicles and great vesicles; multivesicular bodies, endosomes and lysosomes mainly viewed on supranuclear and apical positions. Presence of an appocrine secretory pathway was characterized in P cells through the occurrence of apical cytoplasmic expansions, protruding into the ducts epididymidis luminal compartment.

Morphological aspect of the midgut of anopheles aquasalis (Curry, 1932) Insecta: Diptera / Aspecto morfológico del intestino medio de anopheles aquasalis (Curry, 1932) Insecta: Diptera

The midgut of adult female Anopheles aquasalis presents a narrow anterior or thoracic region and a distensible posterior or abdominal region constituted by the epithelium formed by a cell layer whose apical portion presents microvilli and the basal portion, a basal labyrinth. The thoracic region revealed heterogeneous cellular staining affinity mainly by the presence of acidic components. The ultrastructural aspect showed columnar cells with the presence of the vesicle, mitochondria, endoplasmic reticulum and secreting cells. The abdominal region of the midgut revealed an irregular epithelium whose cells presented a basophilic cytoplasm and acidophil granules. It was also found secreting and/or basal cells with narrow cytoplasm. The ultrastructural observation of this region demonstrated cells with evident nucleus, endoplasmic reticulum and mitochondria. Larger vesicles and small granules were found distributed throughout the cytoplasm. The basal lamina that supports the epithelium presented a generally irregular aspect and the muscle fibers have longitudinal and circular organization and were found separating the epithelium from the haemocel. This study will contribute to analyses on the vector mosquito-parasite interaction mechanism in this specimen.

La seccion media del intestino de la hembra de Anopheles aquasalis presenta una estrecha region anterior o toráxica y una region posterior o abdominal constituida por el epitelio formado por una camada de células cuya porcion apical presenta microvilosidades y la porcion basal presenta un laberinto basal. La region toráxica reveló afinidad de tintura celular principalmente para componentes acídicos. El aspecto ultra estructural mostró células columnares con la presencia de la vesícula, mitocondrias, retículo endoplasmático y células secretoras. La region abdominal del intestino medio reveló un epitelio irregular con células con citoplasma basófilo y granulos acidófilos. También se encontraron células secretoras y/o básales con citoplasma estrecho. La observacion ultra estructural de la region mostró células con núcleos, retículo endoplasmático y mitocondrias evidentes. Vesículas largas y granulos pequeños fueron encontrados distribuidos por todo el citoplasma. La lámina basal que apoya el epitelio presentó un aspecto irregular y las fibras musculares tienen organizacion longitudinal y circular y separan el epitelio del hemocele. Este estudio contribuirá al análisis del mecanismo de interaccion entre el mosquito y el parásito en este espécimen.

Three-dimensional characteristics of bacteriae in the filiform and fungiform papillae of rat: a TEM and HRSEM study / Características tridimensionales de bacterias en las papilas filiformesy fungiformes de rata: un estudio en MET y ARMEB

Tongue mucosae of one day postnatal rat was examined by transmission electron microscopic and HRSEMmethods.The specimens were fixed using Karnovsky solution and embedded in Spurr resin for transmission electron microscopy. For HRSEM methods, the samples were fixed in 2 percent osmiun tetroxide, dehydrated in alcohol, critical point dried and coated with gold-palladium. The results demonstrated that the surface of tongue present the filiform and fungiform papillae covered by numerous keratinized epithelial cells. The bacteriae are attached to the surfaces of microplicae at random, demonstrating in their three-dimensional HRSEM images. At high magnification, the transmission electron microscopic images revealed the adhesion of bacteriae to the cell membrane by glycocalyx. The fibrillar structures surrounding the bacteriae are clearly seen.

Un día después del nacimiento, la mucosa de la lengua una rata fue examinada por el microscopio electrónico de transmisión y método de ARMEB. Los especímenes fueron fijados mediante uan solución Karnovsky y embebido en resina Spurr para microscopía electrónica de transmisión. Para el método ARMEB, las muestras fueron fijadas en tetróxido de osmio 2 por ciento, deshidratados en alcohol, secados al punto crítico y recubierto con oro-paladio. Los resultados demostraron que la superficie de la lengua presentaba papilas filiformes y fungiformes cubiertas por numerosas células epiteliales queratinizadas. Las bacterias se unen a las superficies de las microplicas al azar, lo que se demuestra en sus tres dimensiones las imágenes en ARMEB. A gran aumento, las imágenes del microscopio electrónico de transmisión revelan la adhesión de bacterias a la membrana celular por el glicocalix. Las estructuras fibrilares que rodean a las bacterias son claramente visibles.

Spherites in the midgut epithelial cells of the sugarcane borer parasitized by Cotesia flavipes

Diatraea saccharalis, the main pest of sugarcane, has been controlled by Cotesia flavipes. Very little is known about the effect of parasitism on the host organs, including the midgut. The Lepidoptera midgut epithelium is composed of columnar, goblet, regenerative, and endocrine cells. Spherites have been described in columnar and regenerative cells of several Lepidoptera species, and presented a lot of functional meaning. We identified spherites in the midgut epithelial cells of non-parasitized D. saccharalis larvae analyzed the effect of parasitism on spherite morphology and distribution along the length of the midgut. Midgut fragments of both non-parasitized and parasitized larvae were processed for transmission electron microscopy. All the midgut epithelial cells showed spherites, but they were not preferentially located in a particular part of the cells. Parasitized larvae had more spherites, mainly in the columnar cells, than non-parasitized larvae. This observation was associated with an ionic imbalance within the insect host. Spherites were more abundant in the anterior midgut region than in other regions, which suggests that this region is involved in ion transport by intracellular and/or paracellular route.The morphological variability of spherites in the cells of parasitized larvae was related to the developmental stages of these structures.

Las mitocondrias en la transformación celular del epitelio mamario / The mitochondria in the mammary epithelial cells transformation

Antecedentes: La adquisición del fenotipo metastático es el resultado de la potente acción generada por oncogenes transformantes sobre una célula normal los cuales con la consiguiente expresión de oncoproteínas determinan drásticos cambios tanto en la morfología como en los volúmenes de los componentes celulares, generando una célula con diferente funcionalidad. Este mecanismo corresponde a la transformación celular. Objetivo: Precisar las modificaciones que caracterizan al mecanismo transformante en células de epitelio mamario transfectado con el oncogén ras (HC11 ras) en comparación con su tipo celular normal (HC11GM). Método: Se estudió con microscopia electrónica de transmisión aplicando técnicas morfométricas a estos tipos celulares con énfasis en las mitocondrias, cuantificando variaciones del organelo generador de energía. Resultados: Todos los parámetros mitocondriales evaluados en el tipo celular transformado presentan diferencias significativas con respecto a la célula normal. Conclusión: Las drásticas modificaciones experimentadas por las mitocondrias se reflejan en la adquisición de nuevos requerimientos energéticos y metabólicos en la célula transformada.

Background: The acquisition of the metastatic phenotype is the result of the transforming oncogene powerful action over a normal cell which with the subsequent oncoprotein expression leads to drastic changes in morphology as well as in cell components volumes, generating a cell with a different function. This mechanism relates to the cell transformation. Objective: To specify the modifications that characterize the transforming mechanism in mammary epithelial cells transfected with the ras oncogene comparing them with its normal cell type. Method: Transmission electronic microscopy using morphometric techniques was applied to this cell types, emphasizing mitochondria variations, trying to clarify its role in each cell type metabolism. Results: Everyone mitochondrial parameters examined in transformed cell type present significant differences regarding to the normal cells. Conclusión: The drastic changes in mitochondria are reflected in the acquisition of new energy requirements and metabolism in the transformed cell.

Análise ultraestrutural e de fatores de crescimento de diferentes métodos de preservação da membrana amniótica utilizada em cirurgia ocular / Ultrastructural and growth factor analysis of amniotic membrane preserved by different methods for ocular surger

OBJETIVO: Comparar, por microscopia eletrônica, a integridade anatômica e a presença de fatores de crescimento e citocinas da membrana amniótica preservada com glicerol/MEM (1:1) e dimetilsulfóxido puro. MÉTODOS: As membranas amnióticas preservadas em glicerol/MEM (1:1) ou dimetilsulfóxido puro foram processadas para microscopia eletrônica de transmissão e varredura. Como controle, membrana amniótica fresca foi imediatamente fixada após coleta e processada para microscopia eletrônica. As citocinas e os fatores de crescimento avaliados foram: TGF-beta- fator transformador de crescimento beta; TGF-beta ativ- fator transformador de crescimento beta ativado; EGF- fator recombinante de crescimento epitelial humano; FGF-4- fator de crescimento fibroblástico 4; FGF-beta- fator de crescimento fibroblástico básico; IL-4- interleucina 4; PGE2- prostaglandina E2; IL-10- interleucina 10; KGF- fator de crescimento de queratinócito; HGF- fator de crescimento de hepatócito. RESULTADOS: As membranas amnióticas do grupo controle apresentavam epitélio íntegro, com microvilos na superfície e complexos juncionais entre as células e a membrana basal. As membranas amnióticas preservadas em glicerol/MEM tinham aspecto semelhante às do controle, com maior altura das células epiteliais. Já as membranas amnióticas preservadas em dimetilsulfóxido mostraram redução das junções intercelulares e destacamento do epitélio da membrana basal. As citocinas e fatores de crescimento não apresentaram diferenças entre os grupos, exceto FGF-4, FGF-beta, PGE2 e KGF. CONCLUSÕES: A membrana amniótica preservada em meio glicerol/MEM apresentou melhor integridade tecidual, com menor desprendimento do epitélio da membrana basal, em comparação com a preservada no dimetilsulfóxido puro. Os fatores de crescimento e citocinas estavam, em sua maior parte, preservados com as duas técnicas de preservação.

PURPOSE: To compare the anatomical structure and the presence of growth factors and cytokines of amniotic membrane preserved in glycerol/MEM (1:1) or undiluted dimethyl sulfoxide through electron microscopy. METHODS: Amniotic membrane preserved in glycerol/MEM (1:1) or undiluted dimethyl sulfoxide were processed for transmission and scaning electron microscopy. As control, freshly collected amniotic membrane was fixed and processed for electron microscopy. The cytokines and growth factors assessed were: TGF-beta (transforming growth factor beta); TGF-b activ (activated transforming growth factor beta); EGF (epidermal growth factor); FGF-4 (fibroblast growth factor 4); bFGF (basic fibroblast growth factor); IL-4 (interleukin 4); PGE2 (prostaglandin E2); IL-10 (interleukin 10); KGF (keratinocyte growth factor); HGF (hepatocyte growth factor). RESULTS: Amniotic membrane from the control group showed intact epithelium, with surface microvilli and junctional complexes between the cells and the basal membrane. Glycerol/MEM preserved amniotic membrane had similar aspect to the control, with higher epithelial cells. Those amniotic membranes preserved in dimethyl sulfoxide disclosed less intercellular junction and detachment of the epithelium from the basal membrane. The cytokines and growth factors did not disclose significant differences, except for FGF-4, bFGF, PGE2 and KGF. CONCLUSIONS: Amniotic membrane preserved in glycerol/MEM showed a better tissue structure, with less detachment of the epithelium from the basal membrane, in comparison to undiluted dimethyl sulfoxide. The majority of the growth factors and cytokines were kept with both techniques of preservation.